Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ER(T) and Cre-ER(T2) recombinases.
Fiche publication
Date publication
novembre 1999
Journal
Nucleic acids research
Auteurs
Membres identifiés du Cancéropôle Est :
Pr CHAMBON Pierre, Dr METZGER Daniel
Tous les auteurs :
Indra AK, Warot X, Brocard J, Bornert JM, Xiao JH, Chambon P, Metzger D
Lien Pubmed
Résumé
Conditional DNA excision between two LoxP sites can be achieved in the mouse using Cre-ER(T), a fusion protein between a mutated ligand binding domain of the human estrogen receptor (ER) and the Cre recombinase, the activity of which can be induced by 4-hydroxy-tamoxifen (OHT), but not natural ER ligands. We have recently characterized a new ligand-dependent recombinase, Cre-ER(T2), which was approximately 4-fold more efficiently induced by OHT than Cre-ER(T) in cultured cells. In order to compare the in vivo efficiency of these two ligand-inducible recombinases to generate temporally-controlled somatic mutations, we have engineered transgenic mice expressing a LoxP-flanked (floxed) transgene reporter and either Cre-ER(T) or Cre-ER(T2) under the control of the bovine keratin 5 promoter that is specifically active in the epidermis basal cell layer. No background recombinase activity could be detected, while recombination was induced in basal keratinocytes upon OHT administration. Interestingly, a dose-response study showed that Cre-ER(T2) was approximately 10-fold more sensitive to OHT induction than Cre-ER(T).
Mots clés
Animals, Enzyme Induction, Epidermis, drug effects, Estrogen Receptor Modulators, pharmacology, Genes, Reporter, Humans, Integrases, biosynthesis, Keratinocytes, drug effects, Mice, Mice, Transgenic, Mutagenesis, Site-Directed, Receptors, Estrogen, antagonists & inhibitors, Recombinant Fusion Proteins, drug effects, Tamoxifen, analogs & derivatives, Viral Proteins
Référence
Nucleic Acids Res.. 1999 Nov;27(22):4324-7